Quantitative research of multiple active components in Fukejing Capsule based on ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry / 中草药

ABSTRACT

Objective To establish a quantitative analysis method of multiple active components (gallic acid, tanshinol, ferulic acid, rosmarinic acid, salvianolic acid B, luteolin, apigenin, aloe-emodin, emodin, butylidenephthalide, and levistilide A) in Fukejing Capsule (FC) based on ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UPLC-Q- Orbitrap HRMS). Methods The column was BEH C18 (50 mm × 2.1 mm, 1.7 μm) and the mobile phase was consisted of acetonitrile-water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min with gradient elution; Mass spectrometer conditions Heated electrospray ionization source (HESI) and full mass quantify methods were used to perform the determination, and the results of the contents were imported into the muti-data processing soft ware SIMCA14.0 to make a quality evaluation. Results Under the optimized conditions, gallic acid, tanshinol, ferulic acid, rosmarinic acid, salvianolic acid B, luteolin, apigenin, aloe-emodin, emodin, butylidenephthalide, and levistilide A all showed good liner relationship (r ≥ 0.999 5) in the range of 0.15—1.50, 0.80—8.00, 1.50—15.00, 0.04—0.40, 0.015—0.150, 0.10—1.00, 0.004—0.040, 0.004—0.040, 0.02—0.20, 0.01—0.10, and 0.04—0.40 μg/mL, respectively; The results of the accuracy, the repeatability and the stability all reached the standards (RSD ≤ 4%); The recoveries ranged from 98%—102% and RSDs were below 3%; The result of the content ranges in different batches were 26.36—31.03 μg/g (gallic acid), 178.85—210.79 μg/g (tanshinol), 320.91—343.16 μg/g (ferulic acid), 2.84—3.09 μg/g (rosmarinic acid), 3.55—4.25 μg/g (salvianolic acid B), 27.33—32.36 μg/g (luteolin), 1.89—2.13 μg/g (apigenin) A, 0.47—0.60 μg/g (aloe-emodin), 3.17—3.57 μg/g (emodin), 1.99—2.54 μg/g (butylidenephthalide), and 7.51—8.53 μg/g (levistilide A). The analysis results showed that the quality of the most batches was stable, the tanshinol and ferulic acid had a great influence on the quality of the medicine, and could be monitored to ensure the quality of different batches. Conclusion The methods established in this paper have a high sensitivity and accuracy; The results of the methodology conform to the relevant requirements and the methods can rapidly determinate the multiple active components in FC; The research also provides a new scientific basis and reference for the quality assessment at the same time.